Effects Of Danxingtobgluo Decoction On The Expression Of Nerve Growth Factor、matrix Metalloproteinases-2in Cerebral Ischemia Reperfusion Injury In Rats

Objective:Cerebrovascular disease(CVD) is the variety of vasogenic brain lesionscaused by brain dysfunction.Stroke is acute cerebral circulation disorders quickly leadto localized or diffuse brain impairment in clinical events.CVD is a common andfrequently-occurring disease of the nervous system, the mortality rate is about10%ofall diseases.and it is one of the three major causes of death of the current threats tohuman health.50%to70%of the survivors who left paralysis, aphasia and severedisability impose a heavy burden to the state, society and family.In the cerebrovasculardisease, ischemic cerebrovascular disease and the incidence of hemorrhagiccerebrovascular disease in the ratio of about3:1.It has so high morbidity, mortality thatclinical focus on prevention and treatment of the diseases.The effective treatment ofcerebral infarction is undoubtedly the early restoration of blood supply.Recovery ofoxygen and glucose supply and cerebral metabolism in the infarcted area after bloodflow reperfusion,the brain tissue damage should be restored.In practice, however not thecase,because of the valid time,it is reperfusion time window.Cerebral ischemiaultra-early treatment time window is in6hours.If the reperfusion pass over this windowtime limit,brain injury will intensify, resulting in reperfusion injury,leading tointracranial hemorrhage, cerebral edema and other serious complications.Currentlyconsidered,the mechanisms of reperfusion injury are:formation of free radicals and the"waterfall" chain reaction,calcium overload in nerve cells,cell toxicity of excitatoryamino acid and so on,which lead to nerve cell damage.The core to reduce reperfusioninjury is actively taking brain protection measures.Studies have shown that cerebral edema and inflammation of ischemic areas caused by leukocyte infiltration is important mechanism of reperfusion injury,and theincreased expression of MMP-2is one of the reasons.At the same time,cerebralischemia can induce the expression of nerve growth factor,endogenous and exogenousNGF in both inhibition of apoptosis, and protection of nerve cells.The prescription ofDanxingtongluo decoction was summed up after years of clinical experience,whichSalvia Huoxuetongluo and the brain networks could effectively eliminate phlegm andblood stasis;at the same time, rhubarb, chuanxiong, sophora japonica, panaxnotoginseng, safflower Tongfu Xiere, Xingqi and blood circulation;accompanied by askullcap, Yuan Participation heat dampness and phlegm.Various drugs played a total theTongfu circulation, expectorant resuscitation efficacy.Previous findings show the role ofDanxingtongluo decoction on the treatment of ischemic stroke,however,the protectiveeffect on cerebral ischemia and reperfusion and of MMP-2, of NGF role is not clear.Inthis study, through observe the expression of MMP-2and NGF protein in the braintissue,after Danxingtongluo decoction treat the rats of cerebral ischemia reperfusion,wecan investigate its protective effect on cerebral ischemia reperfusion injury andmolecular mechanisms,to provide an objective basis for the clinical application ofDanxingtongluo decoction.Methods:40SD rats of either sex, were randomly divided into sham operationgroup, ischemia-reperfusion group, nimodipine control group and Danxingtobgluodecoction pretreatment group of lose、high dose,(n=8).The sham operation group andischemia reperfusion group of saline irrigation to1ml/100g,nimodipine group gavagewith nimodipine suspension (1.0ml/100g),Of Danxingtongluo decoction respectivelyirrigation to the small dose group0.75ml/100g、the1.5ml/100g of high-dose group, allorally once a day, seven days in a row,2h after the last administration to surgery.Usingthe suture-occuluded method in rat middle cerebral artery to make focal cerebralischemia reperfusion model (sham group not treated).After cerebral ischemia2h toreperfusion in, and observe at24h of reperfusion.In accordance with the the Longarating standards, the qualifier to detect the expression levels of MMP-2andNGF-positive cells.Added at any time of failure and death in rats.Results:1. The model is a success rate of68.09%, drug treatment followed by reperfusionfor24hours neurological score was significantly lower than the model group (P <0.05).2. HE rapid routine staining of ultrathin sections of brain tissue after cerebralischemia reperfusion to observe the morphological of tissue:Sham operation group, the cell structure is normal;Cerebral ischemia and reperfusion were seen in the fuzzystructure of nerve cells in the brain tissue,and cell body swelling,varying degrees ofnuclear pyknosis,karyolysis,softening foci formation, also the infiltration ofneutrophils;Nimodipine、Danxingtongluo decoction low and high dose group alsovisible derangement of the nerve cells,cell shrinkage and nuclear pyknosis stainphenomenon, but the extent is more significantly reduced than ischemia-reperfusiongroup,especially Danxingtongluo decoction high dose group.3. Ultra-thin slices of brain tissue by immunohistochemical method to observe theexpression levels of MMP-2、 NGF-positive cells:after cerebral ischemia andreperfusion,treatment groups show that the expression of MMP-2decreasedsignificantly, and NGF expression was significantly increased in the ischemichippocampus.Compared with cerebral ischemia reperfusion group, the difference wasstatistically significant (P <0.05).The effect most significant group is Danxingtongluodecoction high dose(P <0.01).Conclusions:Danxingtongluo decoction has a protective effect on cerebralischemia and reperfusion injury, can effectively improve the symptoms of neurologicaldeficit, and its mechanism may be related to inhibition of MMP-2protein synthesis,increase NGF expression.